Zwitterionic Surfactant Hemolysis Reagent for HbA1c Enzymatic Analysis

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Solution Overview

Problem

Current methods for measuring glycated hemoglobin (HbA1c) using enzymatic methods are limited by slow reaction times and low sensitivity, making them inefficient for accurate and rapid quantification.

Innovation Solution

A hemolysis reagent composition containing a zwitterionic surfactant and a nitrous compound, such as sodium nitrite, is used to accelerate hemolysis and improve reaction sensitivity, allowing for faster and more accurate measurement of HbA1c by facilitating the exposure of the N-terminal β-chain of hemoglobin and enhancing enzymatic reactions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of time

If conventional hemolysis reagents are used for HbA1c measurement, then the measurement can be performed, but the reaction time is long and reaction sensitivity is low

Engineering Contradiction:
Improvemeasurement timeVSAvoidreaction sensitivity
Core Design Contradiction:
Loss of timeVSReliability

Solution Approach 1:

The patent changes the chemical parameters of the hemolysis reagent by introducing a zwitterionic surfactant with specific molecular structure (containing both positive and negative charges) and controlling its concentration (0.1-1.0% w/v), along with adjusting pH (6.0-8.0) and ionic strength, to optimize both hemolysis speed and enzymatic reaction sensitivity for HbA1c measurement

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite hemolysis reagent system combining zwitterionic surfactant with traditional hemolysis agents (such as surfactants, chaotropes, or acids), where the zwitterionic surfactant component specifically enhances the exposure of N-terminal β-chain while the other components provide hemolysis function, achieving synergistic effects that improve both reaction speed and sensitivity

Inventive Principle:
Principle #40Composite materials

2Measurement precision

If standard HbA1c measurement methods are used, then accurate quantification can be achieved, but the operation is complex and requires specialized laboratory equipment

Engineering Contradiction:
ImproveHbA1c quantification accuracyVSAvoidlaboratory equipment requirement
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts and isolates the critical functional component (zwitterionic surfactant) that enables both efficient hemolysis and enhanced enzymatic reaction, allowing the measurement system to be simplified from complex HPLC or immunoassay equipment to a streamlined enzymatic colorimetric assay that can be performed with basic spectrophotometry

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The zwitterionic surfactant acts as an intermediary substance that facilitates the interaction between hemoglobin and the enzymatic reagents, improving the accessibility of the N-terminal β-chain to enzymes and thereby enhancing the overall measurement efficiency without requiring complex separation or purification steps

Inventive Principle:
Principle #24Intermediary (Mediator)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The reagent composition significantly reduces measurement time and improves accuracy by accelerating hemolysis and increasing reaction sensitivity, enabling more precise quantification of HbA1c levels.

Implementation Method 1

accelerating hemolysis and improving reaction sensitivity, allowing for faster and more accurate measurement of HbA1c by facilitating the exposure of the N-terminal β-chain of hemoglobin

Methodology Applied
Scientific EffectHemolysis:

Implementation Method 2

the nitrous compound makes the protein structure of the hemoglobin flexible so that the amino acid sequence of the N-terminal β-chain of hemoglobin can be readily cut off

Methodology Applied
Scientific EffectProteolysis:

Implementation Method 3

producing hydrogen peroxide by reacting fructosyl peptide oxidase (FPOX) with the fructosyl amino acids obtained in step 3

Methodology Applied
Scientific EffectOxidation: Oxidation

Implementation Method 4

oxidizing the hydrogen peroxide produced in step 4 by reacting with peroxidase (POD)

Methodology Applied
Scientific EffectPeroxidase reaction:

Data Source

PatentEP2813854B1Hemolysis reagent composition for hemoglobin a1c quantitative analysis using enzymatic method
Publication Date: 2016.09.07 I SENS INC
  • EP2813854B1 patent drawingFigure 1(a)~1(f)
  • EP2813854B1 patent drawingFigure 2(a)~2(e)
  • EP2813854B1 patent drawingFigure 3(a)~3(f)

AI summary

Disclosed is a hemolysis reagent composition for the quantitative analysis of glycated hemoglobin (HbAlc) via an enzymatic method. The hemolysis reagent compositions of the present invention containing a zwitterionic surfactant and a nitrous compound considerably accelerate hemolysis rate due to the presence of the zwitterionic surfactant, and also induce the N-terminal β-chain of hemoglobin released by hemolysis to be exposed to the outside of the hemoglobin molecule and assists the participation of a greater number of molecules in the reaction, thereby improving reaction sensitivity and reaction rate of the glycated hemoglobin, and the nitrous compound makes the protein structure of the hemoglobin flexibly modified so that the amino acid sequence of the N-terminal β-chain of hemoglobin can be readily cut off, thereby significantly reducing the entire time required for the measurement and improving the accuracy of the measurement.